Zhang Xinjing, Zhang Yuanyuan, Zhang Yu, Liang Feng, Yuan Wanting, Yang Shouxiang, Mei Long, Wang Qingqing, Shao Wei, Sun Xiaoyu
Key Laboratory of Oral Diseases Research of Anhui Province, College & Hospital of Stomatology, Anhui Medical University, Hefei, China.
Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Pathogen Biology, School of Basic Medical Sciences, Anhui Medical University, Hefei, Anhui, China.
J Periodontol. 2025 Aug 17. doi: 10.1002/jper.11394.
Long noncoding RNAs (lncRNAs) are emerging regulators of periodontal inflammation. This study investigates the role of lncRNA MIR100HG in periodontitis and its interaction with the RNA-binding protein Quaking (QKI).
In vitro, human gingival fibroblasts (HGFs) were stimulated with Porphyromonas gingivalis lipopolysaccharide (Pg.LPS, 200 ng/mL, 24 h). In vivo, ligature-induced periodontitis was established in mice. RNA sequencing, RNA immunoprecipitation (RIP), fluorescence in situ hybridization (FISH), and Western blotting were employed to dissect molecular mechanisms.
MIR100HG was downregulated in inflamed gingival tissues (RNA-seq). FISH and Subcellular RNA fractionation analysis confirmed MIR100HG was primarily localized in the nucleus of HGFs. Knockdown of MIR100HG attenuated Pg.LPS-stimulated inflammation (interleukin [IL]-1β, IL-6 and tumor necrosis factor alpha [TNF-α]) in HGFs, while overexpression exacerbated cytokine release. The interaction between QKI and MIR100HG was identified by RNA immunoprecipitation assays. QKI silencing reversed anti-inflammatory effects of MIR100HG knockdown, linking this axis to nuclear factor kappa B (NF-κB) activation. In mice, knockdown of MIR100HG significantly mitigated alveolar bone loss and the levels of inflammatory markers.
This study identifies MIR100HG as a key lncRNA in periodontitis, showing its downregulation in human and murine models. MIR100HG knockdown alleviates periodontal inflammation via a QKI-mediated feedback loop, underscoring its potential as a therapeutic target for periodontitis management.
Long non-coding RNAs (lncRNAs) are emerging as critical regulators in the pathogenesis of periodontitis, yet the functional roles of most differentially expressed lncRNAs remain poorly understood. In this study, we identified MIR100HG as a significantly downregulated lncRNA in periodontitis-affected gingival tissues through RNA transcriptome sequencing. Functional experiments revealed that MIR100HG knockdown attenuates Porphyromonas gingivalis lipopolysaccharide (Pg.LPS) -induced inflammation in human gingival fibroblasts (HGFs), as evidenced by reduced levels of proinflammatory cytokines (interleukin [IL] -1β, IL-6, and tumor necrosis factor [TNF] -α), whereas MIR100HG overexpression exacerbates inflammatory responses. Mechanistically, MIR100HG activates the nuclear factor kappa B (NF-κB) signaling pathway by interacting with the RNA-binding protein Quaking (QKI), a known suppressor of NF-κB. This interaction forms a negative feedback loop that regulates inflammatory responses. Importantly, MIR100HG knockdown alleviated periodontal inflammation and bone loss in periodontitis mouse model, further supporting its pivotal role in disease progression. Our findings demonstrate that MIR100HG modulates periodontal inflammation through a QKI- mediated mechanism, highlighting its potential as a therapeutic target for periodontitis. This study provides novel insights into lncRNA-mediated immune regulation and highlights its potential as a therapeutic target for managing periodontitis.
长链非编码RNA（lncRNA）是牙周炎炎症反应中新兴的调控因子。本研究旨在探讨lncRNA MIR100HG在牙周炎中的作用及其与RNA结合蛋白颤抖蛋白（QKI）的相互作用。
体外实验中，用牙龈卟啉单胞菌脂多糖（Pg.LPS，200 ng/mL，24小时）刺激人牙龈成纤维细胞（HGFs）。体内实验中，在小鼠中建立结扎诱导的牙周炎模型。采用RNA测序、RNA免疫沉淀（RIP）、荧光原位杂交（FISH）和蛋白质免疫印迹法来剖析分子机制。
在炎症牙龈组织中，MIR100HG表达下调（RNA测序）。FISH和亚细胞RNA分级分析证实MIR100HG主要定位于HGFs细胞核中。敲低MIR100HG可减轻Pg.LPS刺激的HGFs炎症反应（白细胞介素[IL]-1β、IL-6和肿瘤坏死因子α [TNF-α]），而过表达则加剧细胞因子释放。通过RNA免疫沉淀实验确定了QKI与MIR100HG之间的相互作用。敲低QKI可逆转敲低MIR100HG的抗炎作用，将该轴与核因子κB（NF-κB）激活联系起来。在小鼠中，敲低MIR100HG可显著减轻牙槽骨丧失和炎症标志物水平。
本研究确定MIR100HG是牙周炎中的关键lncRNA，在人和小鼠模型中均显示其表达下调。敲低MIR100HG可通过QKI介导的反馈环减轻牙周炎炎症，突出了其作为牙周炎治疗靶点的潜力。
长链非编码RNA（lncRNA）正成为牙周炎发病机制中的关键调控因子，但大多数差异表达lncRNA的功能作用仍知之甚少。在本研究中，我们通过RNA转录组测序确定MIR100HG是牙周炎受累牙龈组织中显著下调的lncRNA。功能实验表明，敲低MIR100HG可减轻牙龈卟啉单胞菌脂多糖（Pg.LPS）诱导的人牙龈成纤维细胞（HGFs）炎症，促炎细胞因子（白细胞介素[IL]-1β、IL-6和肿瘤坏死因子[TNF]-α）水平降低证明了这一点，而MIR100HG过表达则加剧炎症反应。从机制上讲，MIR100HG通过与RNA结合蛋白颤抖蛋白（QKI）相互作用激活核因子κB（NF-κB）信号通路，QKI是已知的NF-κB抑制剂。这种相互作用形成负反馈环，调节炎症反应。重要的是，敲低MIR100HG可减轻牙周炎小鼠模型中的牙周炎症和骨丧失，进一步支持其在疾病进展中的关键作用。我们的研究结果表明，MIR100HG通过QKI介导的机制调节牙周炎炎症，突出了其作为牙周炎治疗靶点的潜力。本研究为lncRNA介导的免疫调节提供了新见解，并突出了其作为治疗牙周炎靶点的潜力。
